Structural and Functional Analysis of Disease-Linked p97 ATPase Mutant Complexes
Abstract
IBMPFD/ALS is a genetic disorder caused by a single amino acid mutation on the p97 ATPase, promoting ATPase activity and cofactor dysregulation. The disease mechanism underlying p97 ATPase malfunction remains unclear. To understand how the mutation alters the ATPase regulation, we assembled a full-length p97^(R155H) with its p47 cofactor and first visualized their structures using single-particle cryo-EM. More than one-third of the population was the dodecameric form. Nucleotide presence dissociates the dodecamer into two hexamers for its highly elevated function. The N-domains of the p97^(R155H) mutant all show up configurations in ADP- or ATPγS-bound states. Our functional and structural analyses showed that the p47 binding is likely to impact the p97^(R155H) ATPase activities via changing the conformations of arginine fingers. These functional and structural analyses underline the ATPase dysregulation with the miscommunication between the functional modules of the p97^(R155H).
Additional Information
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Received: 2 July 2021; Accepted: 25 July 2021; Published: 28 July 2021. We thank Ellen Zhong (Massachusetts Institute of Technology) for the brief discussion on the data analysis using cryoDRGN neural networks. We thank Xiaoyi Zhang, Gui Lin, and Daniel Wong for preliminary ATPase activity determination for p97 mutants. We thank Andrey Malyutin for assisting in sample screening at the Beckman Caltech Cryo-EM Center. We acknowledge using the Titan Krios TEM in the Eyring Materials Center (EMC) at Arizona State University (ASU) and funding for the instrumentation from grant number NSF MRI 1531991. We thank the NVIDIA GPU Grant Program to P.-L.C for GPU device support. The project was partially supported by a grant from the National Institute of Neurological Disorders and Stroke (NINDS) (R01NS102279) to T.-F.C. and the ASU startup fund to P.-L.C. Author Contributions: Conceptualization, T.-F.C. and P.-L.C.; investigation, formal analysis, and data curation, P.N., S.L., R.C.A.C., F.W., Y.-P.P., D.R.W. and P.-L.C.; supervision and validation, T.-F.C. and P.-L.C.; writing, P.N., T.-F.C. and P.-L.C. All authors have read and agreed to the published version of the manuscript. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable. Data Availability Statement: Cryo-EM density maps have been deposited in the Electron Microscopy Data Bank (EMDB) under accession numbers EMD-24302 (p97^(R155H)|ATPγS-p47), EMD-24305 (p97^(R155H)-p47), EMD-23191 (p97^(R155H) dodecamer), EMD-24304 (p97^(R155H)|ADP-p47), and EMD-23192 (p97^(R155H) dodecamer II). Model coordinates were deposited in the Protein Data Bank (PDB) under accession numbers 7R7S (p97^(R155H)|ATPγS-p47), 7R7U (p97^(R155H)-p47), 7L5W (p97^(R155H) dodecamer), and 7R7T (p97^(R155H)|ADP-p47). All data are available from the corresponding authors upon request.Attached Files
Published - ijms-22-08079-v2.pdf
Supplemental Material - ijms-22-08079-s001.zip
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Additional details
- PMCID
- PMC8347982
- Eprint ID
- 110339
- Resolver ID
- CaltechAUTHORS:20210820-234106808
- MRI-1531991
- NSF
- NVIDIA Corporation
- R01NS102279
- NIH
- Arizona State University
- Created
-
2021-08-21Created from EPrint's datestamp field
- Updated
-
2021-08-21Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering